PT  - JOURNAL ARTICLE
AU  - Harjung, Alexander
AU  - Fracassi, Alessandro
AU  - Devaraj, Neal
TI  - Encoding extracellular modification of artificial cell membranes using engineered self-translocating proteins
AID  - 10.1101/2023.10.06.561148
DP  - 2023 Jan 01
TA  - bioRxiv
PG  - 2023.10.06.561148
4099  - http://biorxiv.org/content/early/2023/10/06/2023.10.06.561148.short
4100  - http://biorxiv.org/content/early/2023/10/06/2023.10.06.561148.full
AB  - A common method of generating artificial cells is to encapsulate protein expression systems within lipid vesicles. However, to communicate with the external environment, protein translocation across lipid membranes must take place. In living cells, protein transport across membranes is achieved with the aid of complex translocase systems which are difficult to reconstitute into artificial cells. Thus, there is need for simple mechanisms by which proteins can be encoded and expressed inside synthetic compartments yet still be externally displayed. Here we present a genetically encodable membrane functionalization system based on mutants of pore-forming proteins. We show that the membrane translocating loop of α-hemolysin can be engineered to translocate functional peptides up to 52 amino acids across lipid membranes. Engineered hemolysins can be used for genetically programming artificial cells to display interacting peptide pairs, enabling their assembly into artificial tissue-like structures capable of signal transduction.Competing Interest StatementThe authors have declared no competing interest.